Erythromelalgia is a neurovascular disorder which causes pain, swelling, erythema, and warmth of the distal extremities. Primary disease is due to a genetic mutation in the gene, but secondary erythromelalgia can be the consequence of a variety of underlying etiologies, including drug and toxin exposures. The disease is rare, occurring in only 1.3 out of every 100,000 in the United States, and symptoms can vary significantly in severity and presentation. Therefore, it can be difficult to recognize the disorder, identify the source, and promptly treat the condition. We report a reversible cause of erythromelalgia induced by the use of oral cyclosporine. This correlation is poorly documented in literature, with limited accounts identifying an association between erythromelalgia and cyclosporine. As drug-induced erythromelalgia represents a reversible cause of disease, physicians should obtain a detailed medication history during the diagnostic workup, specifically inquiring about the use of cyclosporine.

O crescimento gengival induzido por ciclosporina-A (CG) é caracterizado por uma variedade de sinalizações que envolvem fatores de crescimento e proteoglicanos, porém pouco compreendidas. Investigamos a expressão gênica dos proteoheparans sulfato de superfície celular sindecam-2 (SDC-2), -4 (SDC-4) e betaglicam nesse CG. A quantidade total e relativa de glicosaminoglicanos sulfatados (GAGs) e a distribuição de SDC-2 e SDC-4 no tecido gengival também foram analisadas. Métodos: A expressão de mRNA dos proteoglicanos SDC-2, SDC-4 e betaglicam foi analisada pela reação de polimerase em cadeia e transcrição reversa (RT-PCR) em amostras gengivais de 9 indivíduos com CG (grupo CsA) e 6 com gengiva normal (grupo controle). Os GAGs foram extraídos e purificados dos tecidos gengivais e analisados tanto por eletroforese em gel de agarose quanto por espectrofotometria. Foi realizada uma avaliação imunohistoquímica dos tecidos com anticorpos para SDC-2 e SDC-4, para sua localização nos tecidos. Os grupos foram comparados pelo teste t de Student. Resultados: Todos os proteoglicanos estudados mostraramse aumentados no grupo CsA (165% para SDC-2, 308% para SDC-4 e 42% para betaglicam), comparativamente ao grupo controle (P < 0.0001). Não foram observadas diferenças significativas na quantidade total e relativa de GAGs. A imunohistoquímica mostrou uma distribuição marcante de SDC-2 e SDC-4 no componentes epitelial, conjuntivo, vascular, nervoso e inflamatório, incluindo os compartimentos celulares e matriciais de toda a casuística. Nossos resultados revelam expressão aumentada de mRNA de SDC-2, SDC-4 e betaglicam no crescimento gengival induzido pela ciclosporina-A, porém não se observaram diferenças na quantidade de glicosaminoglicanos sulfatados em relação ao tecido gengival não exposto ao fármaco. / Cyclosporin-induced gingival overgrowth (CIGO) comprises a variety of signaling pathways including growth factors and proteoglycans that remains not fully understood. We investigated the gene expression of the cell-surface heparan sulfate proteoglycans syndecan-2 (SDC-2), -4 (SDC-4) and betaglycan in CIGO. Total and relative amounts of sulfated glycosaminoglycans (GAGs) and the distribution of SDC-2 and SDC-4 in the gingival tissue were also analyzed. Methods: mRNA expression of the proteoglycans SDC-2, -4 and betaglycan was analyzed by reverse transcription polymerase chain reaction (RT-PCR) in gingival samples obtained from 9 individuals with CIGO and 6 with a normal gingiva (control group). GAGs were extracted and purified from gingival tissues and analyzed by agarose gel electrophoresis and spectrophotometry. An immunohistochemical evaluation using panels of antibodies for SDC-2 and -4 was performed for localization in the tissues. Groups were compared by the Student's t test. Results: All proteoglycans expressions revealed increase in the CIGO group (165% for SDC-2, 308% for SDC-4 and 42% for betaglycan) compared to the control group (P < 0.0001). No significant differences were observed for the total and relative amounts of GAGs. Immunohistochemistry showed a marked distribution of SDC-2 and SDC-4 in gingival epithelial, connective, vascular, neural and inflammatory components comprising cellular and matrix environments in both groups. Our results reveal increased mRNA expression of SDC-2, SDC-4 and betaglycan in CIGO, but no significant differences in the sulfated glycosaminoglycans component in situ.

L'ischémie-reperfusion (IR) rénale entraîne des lésions de nécrose tubulaire aigue, nécessitant parfois une épuration extra rénale transitoire voir définitive. La mitochondrie joue un rôle important dans la physiopathologie de ces lésions d'IR en entrainant la mort cellulaire. L'étude de l'IR dans la cellule cardiaque a permis de mettre en évidence le rôle central du pore de transition de perméabilité mitochondriale (mPTP) dans le déclenchement de cette mort cellulaire. La ciclosporine (CsA) a été proposée comme thérapeutique pour protéger la cellule des lésions d'IR en retardant l'ouverture de ce mPTP. Cependant la CsA a des effets rénaux vasoconstricteurs aigus, nécessitant une validation expérimentale de sa protection dans l'IR rénale. Au cours de ce travail de thèse, nous avons mis au point un modèle murin d'IR rénale. Ensuite nous avons montré que le post-conditionnement à la CsA, ainsi que le post-conditionnement ischémique, permettent d'améliorer la fonction rénale avec un retard à l'ouverture du mPTP. Dans un deuxième travail, nous montrons que le pré conditionnement à la CsA est dose et temps dépendant, et médié en partie par l'augmentation d'expression d'une protéine chaperonne, l'Heat Shock Protéine 70 (HSP70). L'injection en bolus de CsA permet également d'améliorer la fonction rénale dans ce modèle d'IR avec un retard à l'ouverture du mPTP. Nos résultats ouvrent de nouvelles perspectives dans la protection rénale, notamment dans la réduction des épisodes d'insuffisance rénale aigue après chirurgie aortique ou en transplantation rénale / Ischemia-reperfusion (IR) is a situation encountered in transplantation or during aortic surgery, which can result in renal damages, requiring sometimes transient or definitive dialysis. Mitochondria play a crucial role in the pathophysiology of IR causing cell death. Previous studies of cardiac IR highlighted the role of mitochondrial permeability transition pore (mPTP). Cyclosporin A (CsA) has been proposed as a treatment to protect the kidney from IR by the delay of the opening of the mPTP. However, CsA has acute renal hemodynamic effects and a long-term toxicity, requiring an experimental validation of its protection in the renal IR. In this work, we developed a mouse model of renal IR. In a first study, we showed that the post-conditioning with CsA and ischemic postconditioning improve renal function with a delay of the opening of the mPTP. In a second study, we showed that a high dose of CsA injected just before the ischemia improves renal function and leads to the delay of the opening of mPTP mediated by an increase of HSP70. Our results open new perspectives in renal protection, especially for reducing episodes of acute renal failure in aortic surgery or in renal transplantation

O crescimento gengival induzido por ciclosporina-A (CG) é caracterizado por uma variedade de sinalizações que envolvem fatores de crescimento e proteoglicanos, porém pouco compreendidas. Investigamos a expressão gênica dos proteoheparans sulfato de superfície celular sindecam-2 (SDC-2), -4 (SDC-4) e betaglicam nesse CG. A quantidade total e relativa de glicosaminoglicanos sulfatados (GAGs) e a distribuição de SDC-2 e SDC-4 no tecido gengival também foram analisadas. Métodos: A expressão de mRNA dos proteoglicanos SDC-2, SDC-4 e betaglicam foi analisada pela reação de polimerase em cadeia e transcrição reversa (RT-PCR) em amostras gengivais de 9 indivíduos com CG (grupo CsA) e 6 com gengiva normal (grupo controle). Os GAGs foram extraídos e purificados dos tecidos gengivais e analisados tanto por eletroforese em gel de agarose quanto por espectrofotometria. Foi realizada uma avaliação imunohistoquímica dos tecidos com anticorpos para SDC-2 e SDC-4, para sua localização nos tecidos. Os grupos foram comparados pelo teste t de Student. Resultados: Todos os proteoglicanos estudados mostraramse aumentados no grupo CsA (165% para SDC-2, 308% para SDC-4 e 42% para betaglicam), comparativamente ao grupo controle (P < 0.0001). Não foram observadas diferenças significativas na quantidade total e relativa de GAGs. A imunohistoquímica mostrou uma distribuição marcante de SDC-2 e SDC-4 no componentes epitelial, conjuntivo, vascular, nervoso e inflamatório, incluindo os compartimentos celulares e matriciais de toda a casuística. Nossos resultados revelam expressão aumentada de mRNA de SDC-2, SDC-4 e betaglicam no crescimento gengival induzido pela ciclosporina-A, porém não se observaram diferenças na quantidade de glicosaminoglicanos sulfatados em relação ao tecido gengival não exposto ao fármaco. / Cyclosporin-induced gingival overgrowth (CIGO) comprises a variety of signaling pathways including growth factors and proteoglycans that remains not fully understood. We investigated the gene expression of the cell-surface heparan sulfate proteoglycans syndecan-2 (SDC-2), -4 (SDC-4) and betaglycan in CIGO. Total and relative amounts of sulfated glycosaminoglycans (GAGs) and the distribution of SDC-2 and SDC-4 in the gingival tissue were also analyzed. Methods: mRNA expression of the proteoglycans SDC-2, -4 and betaglycan was analyzed by reverse transcription polymerase chain reaction (RT-PCR) in gingival samples obtained from 9 individuals with CIGO and 6 with a normal gingiva (control group). GAGs were extracted and purified from gingival tissues and analyzed by agarose gel electrophoresis and spectrophotometry. An immunohistochemical evaluation using panels of antibodies for SDC-2 and -4 was performed for localization in the tissues. Groups were compared by the Student's t test. Results: All proteoglycans expressions revealed increase in the CIGO group (165% for SDC-2, 308% for SDC-4 and 42% for betaglycan) compared to the control group (P < 0.0001). No significant differences were observed for the total and relative amounts of GAGs. Immunohistochemistry showed a marked distribution of SDC-2 and SDC-4 in gingival epithelial, connective, vascular, neural and inflammatory components comprising cellular and matrix environments in both groups. Our results reveal increased mRNA expression of SDC-2, SDC-4 and betaglycan in CIGO, but no significant differences in the sulfated glycosaminoglycans component in situ.

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Ce travail de thèse reposait sur l’hypothèse que la variabilité génétique des protéines « cibles » des médicaments immunosuppresseurs de la famille des inhibiteurs de la calcineurine (ICN ; ciclosporine et tacrolimus) pourrait expliquer une partie de la variabilité observée dans leur efficacité et toxicité. Une revue de la littérature nous a permis de lister un panel de variants génétiques au sein de la voie de la calcineurine, considérés comme étant de bons candidats pour des études en transplantation. Ces variants n’ont pas été associés au risque de rejet aigu ou d’infection grave dans une étude incluant 381 patients transplantés rénaux suivis durant un an après la transplantation. La variabilité pharmacodynamique des ICN a ensuite été explorée au travers des régulations épigénétiques. Une analyse de la méthylation de l’ADN après exposition médicamenteuse a été menée sur deux modèles. Premièrement, la lignée cellulaire JURKAT a été utilisée pour développer la méthode d’immunoprécipitation de l’ADN méthylé (MeDIP). Chez des souris traitées par ciclosporine et tacrolimus durant 3 mois, nous avons ensuite isolé les cellules cibles des médicaments, les lymphocytes T CD4 puis, après immunoprécipitation de l’ADN méthylé et analyse par séquençage pangénomique haut débit (MeDIP-seq, séquençeur Ion Proton), nous avons recherché les régions du génome présentant des différences de méthylation induites par le traitement. L’analyse différentielle bio-informatique a été menée à l’aide des outils SAMtools (Li et col., 2009), BEDtools (Quinlan and Hall, 2010), MACS2 (Zhang et col., 2008) et Diffbind (Stark and Brown, 2011 - Bioconductor). Sur l’ensemble du génome, nous n’avons identifié que 24 régions présentant un niveau de méthylation modifié par l’exposition au tacrolimus. Le promoteur du gène Calm2, codant pour l’isoforme 2 de la calmoduline, semble être davantage méthylé chez les souris traitées. Ces résultats préliminaires semblent prometteurs pour la découverte de biomarqueurs épigénétiques de la réponse thérapeutique aux immunosuppresseurs. / Inter-individual genetic variation might account for diverse efficacy and toxicity of calcineurin inhibitors (cyclosporin and tacrolimus). In particular, some variants located within genes coding for proteins of the calcineurin pathway can explain part of this variability. In this manuscript, a panel of candidate genes was selected based on bibliographic review and tested in a pharmacogenetics study encompassing 381 renal transplants followed for one year after surgery. None of these candidates was associated with the acute rejection or serious infection risks. Furthermore, the pharmacodynamic variability of these drugs was also investigated, exploring the use of epigenomics profiling as proximal readout of the calcineurin inhibition treatment. In particular, we investigated the impact of drug exposure on DNA methylation in two experimental models. Methylated DNA immunoprecipitation followed by high-throughput sequencing (MeDIP-seq, Ion Proton technology) was deployed in JURKAT cell line, used as in vitro model, and in CD4 T lymphocytes isolated from mice treated with either cyclosporin or tacrolimus for three months. After sequencing, the differentiated methylated regions caused by drug exposure were analyzed. Bioinformatics analyses were performed using SAMtools (Li et al., 2009), BEDtools (Quinlan and Hall, 2010), MACS2 (Zhang et al., 2008) and Diffbind (Stark and Brown, 2011 - Bioconductor). Overall, the genome-wide analysis revealed only 24 regions with a differentiated enrichment in DNA methylation after three month-tacrolimus treatment, indicating a targeted effect of these treatments on a subset of key genes. Of note, CALM2 promoter, coding for the calmodulin isoform 2 protein, showed significant hypermethylation in tacrolimus-treated mice. These preliminary results corroborate the interest in using DNA methylation as promising approach to identify candidate biomarkers for therapeutic drug monitoring in calcineurin inhibitor treatments.

Le récepteur neurokinine 1 (NK1R) est impliqué dans la régulation des réponses immunitaires innées et adaptatives. Cependant, les mécanismes par lesquels le NK1R modulerait ces réponses ne sont pas connus. Chez les cellules T, les voies de la calcineurine et de la mTOR constituent les cibles d’immunosuppresseurs, comme la cyclosporine A (CsA), le tacrolimus et la rapamycine. Ainsi, nous avons voulu déterminer si le NK1R pourrait agir sur ces voies et si le blocage pharmacologique du NK1R avec des antagonistes sélectifs, pourrait augmenter l’action de ces immunosuppresseurs sur l’activation des cellules T. Tout d’abord, nos résultats ont montré que les cellules Jurkat (celules T humaines) exprimaient à la fois le gène du NK1R et de son ligand (les endokinines). Ceci suggère l'existence d'une régulation autocrine tachykinergique de la fonction des cellules T. Cette hypothèse est appuyée par nos données, où nous avons observé que le blocage du NK1R avec des antagonistes spécifiques (L-733,060 et L-703,606) chez les cellules Jurkat, inhibe la production d'IL-2 et diminue l'activation du NFAT (substrat de la calcineurine). De façon intéressante, nous avons montré un effet de combinaison entre les antagonistes du NK1R et les inhibiteurs de la calcineurine (CsA et tacrolimus) sur la production d’IL-2 et l’activation du NFAT. En revanche, le blocage du NK1R n'a pas d'effet inhibiteur sur l’activation de la mTOR et la p70S6K, mais réduit la phosphorylation de S6R (Ser235/236) et Akt (Ser473). Enfin, nous n’avons observé aucun effet de combinaison avec la rapamycine et l’antagoniste NK1R sur l’activation de mTOR et de sa voie de signalisation. L’ensemble de nos résultats, démontrent la présence d'un nouveau mécanisme de régulation de NFAT impliquant le système tachykinergique NK1R/endokinines chez les cellules T. Par conséquent, nous suggérons que la combinaison des antagonistes NK1R avec les inhibiteurs de la calcineurine pourrait être une alternative thérapeutique intéressante afin de réduire les doses de CsA et le FK506 dans les protocoles de prévention de rejet de greffes. / There are increasing evidences for a role of the neurokinin 1 receptor (NK1R) in the regulation of innate and adaptive immune systems. However, whether NK1R regulates calcineurin/nuclear factor of activated T cell (NFAT) and mTOR pathways in T cells is unknown. These signalling pathways being targets of the immunosuppressive drugs cyclosporine A (CsA), FK506 and rapamycin respectively. We also examined whether pharmacological blockade of NK1R may be combined to those immunosuppressors to repress T cell activation. In this article, we first show that Jurkat T cells express both genes for NK1R and its ligands endokinins which suggests the existence of an autocrine tachykinergic regulation of T cells function. This hypothesis is supported by our data showing that blockade of this receptor with specific NK1R antagonists inhibits IL-2 production in Jurkat T cells which is associated with the reduction of NFAT activation. Interestingly, we show interplay between NK1R antagonists and calcineurin inhibitors to repress IL-2 production and NFAT activation. In contrast, blockade of NK1R has no inhibitory effect on mTOR and p70S6K activation but reduce S6R (Ser235/236) and Akt (Ser473) phosphorylation. However, combining rapamycin with NK1R antagonist has no enhancing effect on rapamycin-reduced mTOR activation and its signalling pathway. Our findings provide the evidence of a novel mechanism of regulation of NFAT activation-induced IL-2 production in T cells involving the tachykinergic system NK1R/endokinins. These observations may offer new application for NK1R antagonists in transplantation immunotherapy in combination with immunosuppressors.

Implication du PTP dans la mort cellulaire β pancréatique L'hyperglycémie, l'hyperfructosémie et l'ischémie-reperfusion sont délétères pour la viabilité cellulaire β pancréatique, jouant un rôle majeur dans la perte de la masse cellulaire β. Le pore de transition de perméabilité mitochondriale (PTP) est un canal mitochondrial impliqué dans le déclenchement de la mort cellulaire. Des données récentes montrent l'implication du PTP et du stress oxydant dans la toxicité induite par l'ischémie-reperfusion sur cardiomyocytes et également dans la glucotoxicité induite sur cellules endothéliales. La première partie de notre étude a visé à étudier l'implication de l'ouverture du PTP dans la mort cellulaire des cellules INS-1 et des îlots pancréatiques humains soumis à de fortes concentrations de glucose et de fructose. Nous démontrons que l'incubation des cellules INS-1 et des îlots pancréatiques humains en présence de 30 mM de glucose ou 2,5 mM de fructose déclenche une ouverture du PTP et induit la mort cellulaire. La metformine et la Cyclosporine A (CsA) préviennent l'ouverture du PTP et la mort cellulaire induite par le glucose et le fructose. La deuxième partie de notre travail montre que l'exposition des INS-1 à une heure de carence en substrat concomitante d'une hypoxie, suivie d'une restauration des conditions basales conduit à l'ouverture du PTP et à une majoration drastique de la mort cellulaire. Ces deux évènements sont totalement prévenus par l'incubation préalable par la CsA et la metformine mais aussi par la N-Acétyl-Cystéine (NAC) ou par l'exposition à une anoxie, soulignant ainsi le rôle fondamental du stress oxydant dans le déclenchement de l'ouverture du PTP et de la mort cellulaire. Nous montrons qu'au cours de l'ischémie-reperfusion simulée, la production de superoxide est bi-phasique : nous décrivons un premier pic de production au cours de la carence en substrat, lié à un flux reverse d'électrons au sein du complexe I de la chaîne respiratoire. Ce premier pic est suivi d'un deuxième pic de production après la restauration du niveau de substrats et d'O2, lié à l'ouverture du PTP. La NAC, l'anoxie ou la metformine préviennent les deux pics de production de superoxide tandis que la CsA prévient seulement le second pic. Enfin, nous montrons que l'hypoxie seule n'induit ni stress oxydant, ni ouverture du PTP ni mortalité cellulaire. L'ensemble de notre travail démontre le rôle central du PTP dans la gluco-fructotoxicité et dans la toxicité induite par l'ischémie-reperfusion sur la cellule β pancréatique. Ainsi, prévenir l'ouverture du PTP peut-être une approche intéressante pour préserver la viabilité cellulaire β. / PTP involvement in β pancreatic cell death Hyperglycemia, hyperfructosemia and ischemia-reperfusion play a major role in the progression of β cell loss in diabetes mellitus. The permeability transition pore (PTP) is a mitochondrial channel involved in cell death. PTP opening and oxidative stress have been shown to be involved in ischemia-reperfusion injury on cardiomyocytes and in hyperglycemia-induced cell death in endothelial cells. In the first part of this work, we have examined the involvement of PTP opening in INS-1 cells and human pancreatic islets cell death induced by high levels of glucose or fructose. We first reported that Metformin and Cyclosporin A (CsA) prevented Ca2+-induced PTP opening in permeabilized and intact INS-1 cells. We then shown that incubation of INS-1 cells and human islets in the presence of 30 mM glucose or 2.5 mM fructose induced PTP opening and led to cell death. Because both Metformin and CsA prevented glucose and fructose induced PTP opening, and hampered glucose and fructose induced cell death, we conclude that PTP opening is involved in high glucose and high fructose induced INS-1 and human islets cell death. We therefore suggest that preventing PTP opening might be a new approach to preserve β cell viability. In the second part of the work, we demonstrate that the incubation of INS-1 cells in the absence of energy substrates in hypoxic condition for 1 hour followed by incubation in normal condition led to PTP opening and to a dramatic increase in cell death. Both events were totally prevented when PTP opening was inhibited by either Cyclosporin A (CsA) or Metformin or when the cells were incubated in the presence of the antioxidant N-acetyl-cystein (NAC), in anoxia, highlighting the implication of oxidative stress is the commitment of PTP opening. Superoxide production increased during the removal of energy substrates, due to reverse electron flux through complex I and again increased when normal energy substrate and O2 were restored, due to PTP opening. NAC, anoxia or Metformin prevented the two phases of oxidative stress, while CsA prevented only the second one. Hypoxia alone did not induce oxidative stress, PTP opening or cell death. Our work demonstrates the implication of PTP opening in ischemia-reperfusion injury and gluco- fructotoxicty in β pancreatic cells. We therefore suggest that preventing PTP opening might be a new approach to preserve β cell viability.

A fim de prevenir a rejeição do órgão transplantado pelo hospedeiro, se faz necessário o uso de drogas imunossupressoras, como a ciclosporina, que pode levar ao aparecimento de efeitos colaterais como hipertensão arterial, nefrotoxicidade, hepatotoxicidade, e hiperplasia gengival medicamentosa (HGM), que pode ser potencializada se bloqueadores de canal de cálcio como a nifedipina forem associados a fim de controlar a pressão arterial. A patogênese da HGM ainda é incerta, entretanto fatores como a presença de cálculo e placa, concentração plasmática da droga, idade e fatores hormonais podem influenciar as características clínicas e o desenvolvimento da HGM. Recentemente, alguns vírus têm sido associados com a HGM. O HPV (Papiloma Vírus Humano) tem sido associado com casos severos de HGM, enquanto que o EBV (Vírus Epstein-Barr) é associado ao aparecimento de desordens linfoproliferativas pós transplante, que se apresentam como HGM. O objetivo deste trabalho foi avaliar a freqüência e o grau da HGM em pacientes transplantados renais (TR), identificar o EBV e HPV na HGM destes pacientes, e correlacionar a HGM, índice de placa, presença de cálculo e presença do EBV e HPV nos pacientes TR. Foram examinados os prontuários de 58 pacientes TR atendidos no CAPEFOUSP, e os dados com relação à medicação imunossupressora em uso e presença ou ausência de HGM foram registrados. Foram contatados 15 pacientes TR, dos quais foram colhidos dados demográficos, história médica, drogas em uso e história dental. No exame intra-oral foram observados o índice de placa, grau da HGM e presença de cálculo. A HGM foi removida e enviada a Disciplina de Patologia Bucal para análise microscópica. Os espécimes removidos foram comparados com um grupo controle composto por 20 casos de hiperplasia gengival inflamatória. Ambos os grupos foram submetidos ao exame de rotina, enfatizando a presença de coilócitos e a análise molecular, com hibridização in situ para o EBV (sondas EBER e Lytic) e HPV (sonda de amplo espectro e tipos 6/11, 16/18 e 31/33 nos casos positivos para a sonda de amplo espectro). 42% dos pacientes apresentaram HGM grau 1, 50% grau 2 e 8% grau 3. Cálculo estava presente em 50% dos pacientes. O índice de placa médio encontrado foi de 72%. Todas as amostras gengivais removidas cirurgicamente apresentaram um quadro histopatológico compatível com HGM. Os coilócitos estavam presentes em 100% dos casos do grupo de estudo e em 80% dos casos do grupo controle. O HPV esteve presente em 20% dos casos do grupo de estudo e em 10% do grupo controle. O EBV estava presente em 100% dos casos do grupo de estudo e em 90% dos casos do grupo controle, para ambas as sondas, entretanto no grupo de estudo foi observada uma expressão maior do EBV, tanto em quantidade de células marcadas, como em áreas mais profundas. Concluímos que a maioria dos pacientes TR apresentou HGM leve a moderada; EBV foi encontrado em todos os pacientes TR, caracterizando uma infecção oportunista, enquanto que o HPV foi encontrado nas mesmas proporções nos pacientes TR e no grupo controle; não foi encontrada correlação entre índice da HGM, índoce de placa, presença de cálculo e presença do EBV e HPV. / In order to prevent graft rejection in organ transplantation, is necessary the use of immunosuppressive drugs, as cyclosporin, that has several side effects, such as high blood pressure, nephrotoxicity, hepatotoxicity and gingival overgrowth (GO), that can be increased if calcium channel blockers, such as nifedipine, are associated in order to control de blood pressure. The pathogenesis of GO is still uncertain, but some factors such as the presence of calculus and plaque, drug plasmatic concentration, age and hormonal factors can influence the clinical aspects and development of GO. Recently some virus have been associated to GO as well. HPV (Human Papilloma Virus) have been associated to severe cases of GO and EBV (Epstein-Barr Virus) have been associated to posttransplantation lymphoproliferative disorders presenting as GO. The aim of this work was to evaluate GO incidence and score in renal transplant patients (RTP), identify EBV and HPV in GO from RTP, and correlate GO, plaque score, presence of calculus, and presence of EBV and HPV in RTP. We reviewed 58 charts from RTP attending to Special Care Dentistry Center (CAPE-FOUSP). Immunosuppressant drugs and presence or absence of GO were registered. 15 RTP were asked to show up in order to be examined. We collected demographic data, medical history, drugs in use and dental history. In intra-oral exam we observed plaque score, GO score and presence of calculus. GO were removed and sent to Oral Pathology Department for microscopic analysis. GO was compared to a control group composed by 20 cases of inflammatory gingival hyperplasia and both groups were submitted to routine exam emphasizing the presence of koilocytes and to molecular analysis with in situ hybridization for EBV (EBER and Lytic probes) and for HPV (wide spectrum probe and 6/11, 16/18, 31/33 types in cases where wide spectrum were positive). 42% of the patients presented GO score 1, 50% score 2 and 8% score 3. Calculus were presented in 50% of the patients. The average of plaque score was 72%.All GO specimens removed had a histopathological exam compatible with drug induced gingival overgrowth. Koilocytes were presented in 100% of study group (SG) and in 80% of control group (CG). HPV were presented in 20% of the SG and in 10% of the CG. EBV was presented in 100% of SG and in 90% of CG, for both probes, but in SG it could be observed in deeper areas of the epithelium and in a more pronounced expression. We concluded that most RTP presented mild to moderate GO, EBV were found in all RTP, characterizing an opportunistic infection, while HPV were found in the same proportions than in the control group and there were no statistical correlation between GO, plaque score, presence of calculus and presence of EBV and HPV.

A fim de prevenir a rejeição do órgão transplantado pelo hospedeiro, se faz necessário o uso de drogas imunossupressoras, como a ciclosporina, que pode levar ao aparecimento de efeitos colaterais como hipertensão arterial, nefrotoxicidade, hepatotoxicidade, e hiperplasia gengival medicamentosa (HGM), que pode ser potencializada se bloqueadores de canal de cálcio como a nifedipina forem associados a fim de controlar a pressão arterial. A patogênese da HGM ainda é incerta, entretanto fatores como a presença de cálculo e placa, concentração plasmática da droga, idade e fatores hormonais podem influenciar as características clínicas e o desenvolvimento da HGM. Recentemente, alguns vírus têm sido associados com a HGM. O HPV (Papiloma Vírus Humano) tem sido associado com casos severos de HGM, enquanto que o EBV (Vírus Epstein-Barr) é associado ao aparecimento de desordens linfoproliferativas pós transplante, que se apresentam como HGM. O objetivo deste trabalho foi avaliar a freqüência e o grau da HGM em pacientes transplantados renais (TR), identificar o EBV e HPV na HGM destes pacientes, e correlacionar a HGM, índice de placa, presença de cálculo e presença do EBV e HPV nos pacientes TR. Foram examinados os prontuários de 58 pacientes TR atendidos no CAPEFOUSP, e os dados com relação à medicação imunossupressora em uso e presença ou ausência de HGM foram registrados. Foram contatados 15 pacientes TR, dos quais foram colhidos dados demográficos, história médica, drogas em uso e história dental. No exame intra-oral foram observados o índice de placa, grau da HGM e presença de cálculo. A HGM foi removida e enviada a Disciplina de Patologia Bucal para análise microscópica. Os espécimes removidos foram comparados com um grupo controle composto por 20 casos de hiperplasia gengival inflamatória. Ambos os grupos foram submetidos ao exame de rotina, enfatizando a presença de coilócitos e a análise molecular, com hibridização in situ para o EBV (sondas EBER e Lytic) e HPV (sonda de amplo espectro e tipos 6/11, 16/18 e 31/33 nos casos positivos para a sonda de amplo espectro). 42% dos pacientes apresentaram HGM grau 1, 50% grau 2 e 8% grau 3. Cálculo estava presente em 50% dos pacientes. O índice de placa médio encontrado foi de 72%. Todas as amostras gengivais removidas cirurgicamente apresentaram um quadro histopatológico compatível com HGM. Os coilócitos estavam presentes em 100% dos casos do grupo de estudo e em 80% dos casos do grupo controle. O HPV esteve presente em 20% dos casos do grupo de estudo e em 10% do grupo controle. O EBV estava presente em 100% dos casos do grupo de estudo e em 90% dos casos do grupo controle, para ambas as sondas, entretanto no grupo de estudo foi observada uma expressão maior do EBV, tanto em quantidade de células marcadas, como em áreas mais profundas. Concluímos que a maioria dos pacientes TR apresentou HGM leve a moderada; EBV foi encontrado em todos os pacientes TR, caracterizando uma infecção oportunista, enquanto que o HPV foi encontrado nas mesmas proporções nos pacientes TR e no grupo controle; não foi encontrada correlação entre índice da HGM, índoce de placa, presença de cálculo e presença do EBV e HPV. / In order to prevent graft rejection in organ transplantation, is necessary the use of immunosuppressive drugs, as cyclosporin, that has several side effects, such as high blood pressure, nephrotoxicity, hepatotoxicity and gingival overgrowth (GO), that can be increased if calcium channel blockers, such as nifedipine, are associated in order to control de blood pressure. The pathogenesis of GO is still uncertain, but some factors such as the presence of calculus and plaque, drug plasmatic concentration, age and hormonal factors can influence the clinical aspects and development of GO. Recently some virus have been associated to GO as well. HPV (Human Papilloma Virus) have been associated to severe cases of GO and EBV (Epstein-Barr Virus) have been associated to posttransplantation lymphoproliferative disorders presenting as GO. The aim of this work was to evaluate GO incidence and score in renal transplant patients (RTP), identify EBV and HPV in GO from RTP, and correlate GO, plaque score, presence of calculus, and presence of EBV and HPV in RTP. We reviewed 58 charts from RTP attending to Special Care Dentistry Center (CAPE-FOUSP). Immunosuppressant drugs and presence or absence of GO were registered. 15 RTP were asked to show up in order to be examined. We collected demographic data, medical history, drugs in use and dental history. In intra-oral exam we observed plaque score, GO score and presence of calculus. GO were removed and sent to Oral Pathology Department for microscopic analysis. GO was compared to a control group composed by 20 cases of inflammatory gingival hyperplasia and both groups were submitted to routine exam emphasizing the presence of koilocytes and to molecular analysis with in situ hybridization for EBV (EBER and Lytic probes) and for HPV (wide spectrum probe and 6/11, 16/18, 31/33 types in cases where wide spectrum were positive). 42% of the patients presented GO score 1, 50% score 2 and 8% score 3. Calculus were presented in 50% of the patients. The average of plaque score was 72%.All GO specimens removed had a histopathological exam compatible with drug induced gingival overgrowth. Koilocytes were presented in 100% of study group (SG) and in 80% of control group (CG). HPV were presented in 20% of the SG and in 10% of the CG. EBV was presented in 100% of SG and in 90% of CG, for both probes, but in SG it could be observed in deeper areas of the epithelium and in a more pronounced expression. We concluded that most RTP presented mild to moderate GO, EBV were found in all RTP, characterizing an opportunistic infection, while HPV were found in the same proportions than in the control group and there were no statistical correlation between GO, plaque score, presence of calculus and presence of EBV and HPV.